70 Ali and Maulood

Int. J. Biosci. 2011

RESEARCH PAPER OPEN ACCESS

The allelopathic effect of different species of grapevine berries

aqueous extracts on some germination parameters of lettuce

(Lactuca sativa L.) seeds.

Kawa Abdulkareem Ali1*, Pakhshan Mustafa Maulood2

1Field Crops Dep., Agricultural College, Salahaddin University, Kurdistan Region, Iraq

2Biology, College of Science, Salahaddin University, Kurdistan Region, Iraq

Received: 07 October 2011 Revised: 14 November 2011 Accepted: 15 November 2011

Key words: Allelopathy, growth parameters.

Abstract

Allelopathic potential of twelve local varieties of grapevine Vitis vinifera fruits: taefi; zareg-dohuok; Baetmuni; Black-

manga; Sarqala-bakrajo; Awelka; Rash-merii; Mala-Hassan; Ashgar-basraha; Kamalii; Toli-mawelian and Shekh-

nuraddin on some germination and growth parameters of lettuce seeds Lactuca sativa has been investigated . The

aqueous extracts caused significant inhibitory effects on germination rate; germination speed; radical length; plumule

length; radical, plumule and total seedlings dry weight. Bioassays indicated that the inhibitory effects were increased

with increasing the aqueous extracts concentration. These results indicated that there is a considerable variation of

allelopathic activity that could be utilized in the field of controlling weed plants.

*Corresponding Author: Kawa Abdulkareem Ali kawa91961@yahoo.com

International Journal of Biosciences (IJB) ISSN: 2220-6655 (Print) 2222-5234 (Online)

Vol. 1, No. 6, p. 70-80, 2011 http://www.innspub.net

71 Ali and Maulood

Int. J. Biosci. 2011

Introduction

Grapevine Vitis vinifera is a deciduous woody tree

which its fruits can be eaten raw or used for jam, juice,

jelly, vinegar, raisins…etc. it is one of the most

important fruit species due to its different uses

according to the oldest record belongs to 3500-2900

years B.c. it was originated in southern turkey an area

that is called Anatolia (Gokbayrak and Soylemezoglu,

2010). United Nations FAO statistical data indicated

that 75.866 Km2 of land are grapevine, from this area

71% is used for wine, while only 27% is used as fresh

fruit and only 2% is used for drying (raisins), in iraq

the area of grapevine or vineyards grow gradually as

official statistical data declares that about 84000

hectare with yearly production of 265000 ton that

concentrated in the rural mountain area of iraqi

Kurdistan (FAO, 2007) and (FAO, 2003).

The phytochemistry of grapevine has been intensively

studied and the numbers of identified compounds were

exponentially increased with the development of

analytical chemistry techniques, where today more

than 500 compounds have been identified (Ali et al.,

2010), in general these compounds were considered as

plants secondary metabolites which have diverse

chemical structures and known to play an important

role in different plants biological processes (Vining,

1990). Allelopathy, the biological phenomenon that

defined as the beneficial or harmful biochemical

interaction between plants (Ali, 2001, and Rice 1984),

it is well known that the key factor of these biochemical

interactions is chemicals that known as allelochemicals

(Einhilling, 1995) most of these allelochemicals are

plants secondary metabolites due to the reason that

these compounds were formed throughout main

metabolic pathways of carbohydrates, fats, and amino

acids (Macias, 2005). In spite of the large number of

allelopathic studies on different plant species, we did

not find any papers on the allelopathic potentiality of

grapevine fruits (berries). The objective of this study

was investigating the allelopathic effect of different

local varieties on some germination and growth

parameters of lettuce (Lactuca sativa L.) seeds that

used as bioassay target species in this study to estimate

the allelopathic potentiality of these fruits as an

allelopathic source that might be used for controlling

different agricultural pests especially weed plants

(Inderjit and Dakshini, 1995).

Materials and methods

1. Plant materials

Fruits (berries) of different varieties of grapevine (Vitis

vinifera L.) were collected from local varieties such as

Taefi; Zarig-dohouk; Baetmuni; Sarqala-bakrajo;

Awelka; Rash-meri; Mala-Hassan; Ashgar-basraha;

Kamalii; Toli-mawelian and Shek-nuraddin from the

fields of Arbil agricultural research center that

allocated about 3km from city-center of Erbil.

2. Grapevine fruits chemical analysis

Collected samples were randomly selected on the basis

of absence of any physical or disease injury, then

samples were subjected to some chemical analysis as

total soluble solids by Erma refractometer with using

pure-water as blank solution that has been left to stand

for a period of time in room temperature then sending

the boundary line to zero by using the adjusting screw

knob for calibration (Hellman, 2004). Test for

Alkaloid, poly phenolic compounds, carbohydrates,

Tannins, Saponnins, and Glycosids were run on the

fruit berries aqueous extracts (Bamandy, 2006), Poly

phenolic compounds were estimated according to

folin-ciocalteu reagent method (Pastrana-bonilla.,

2003 and Butkhup et al., 2010).

3. Preparation of grapevine fruits aqueous extracts

Grapevine fruits (berries) were placed in an electrical

blinder then blended and filtered by Whatman#1filter

paper after that samples were sterilized with bacterial

filter (Millipore 0.45µ). The collecting extracts were

stored in special dark plastic bottles prepared for such

purposes and reserved in deepfreeze at -20°C for

subsequent use (Wang et al., 2011), after that different

concentrations of the extracts (zero, 5%, 10%, and 15%

72 Ali and Maulood

Int. J. Biosci. 2011

water) were prepared by adding distilled water to the

fruit extracts with considering that zero concentration

contains only distilled water that was used as control

treatment.

Bioassay

Plant seeds that was chosen as an indicator for these

study was Lettuce Lactuca sativa L. seeds. Twenty five

seeds were placed between two sheets of filter paper #1

in 9 cm Petri-dishes then each petri-dish treated with

8ml of the studied concentrations of the grapevine

berries extracts, after that petri-dishes were sealed and

placed in growth chamber 20-25°C under humidity

75% and continuous darkness. The recording data were

germination rate, inhibition of germination rate, speed

of germination (seed/ day), radical length (cm), radical

elongation velocity (cm/day), plumule length(cm),

plumule elongation velocity (cm/day), radical dry

weight( mg), plumule dry weight (mg), and seedling

total dry weight (mg), radical and plumule growth

inhibition rate the equations shown below (De-Oliveira

et al., 2008) (Ali and Aziz, 2002) (Norsworthy, 2003)

and (Jian and Lafitte, 2007).

(Eq. 1)

(Eq. 2)

(Eq. 3)

Where N1, N2, N3…N = proportion of seeds which

germinate on days 1, 2, 3 …n the following setup of

equation.

After three days from incubating first data set were

recorded the germination of seeds were recorded by

considering the protruded radicals, while the radical or

plumule elongation velocity were calculated according

to the following equation:

(Eq. 4)

Radical or plumule dry weight were recorded by

placing the samples in an electrical oven 40°c for 72

hours or until weight consistency to record the dry

weight. Radical or plumule growth inhibition was

calculated according to the equation by estimating the

inhibition depending on the dry weight of radical or

plumule.

(Eq. 5)

Statistical analysis

The study consists of two factors; grapevine varieties,

and extract concentrations with three replicates for

each treatment, the analysis of variance for all data

were performed using Statistical Package for the Social

Sciences (SPSS version 18) and excel statistical

programmer, while for mean comparisons duncan test

was used (Weinberg and Abramowitz, 2008) and

(Field, 2005).

Results and discussion

1. The effect of Grapevine varieties on the studies

parameters

Results indicates significant differences between all

studied parameters (P≥0.01) except radical and total

seedling dry weights as shown in Table (1), where

Baetmuni and Sarqala-Bakrajo recorded highest

germination rate 96.667% comparing to Mala-Hassan

variety that scored lowest germination rate 64.583 %.

The inhibition of germination rate were opposite to

was noticed for germination rate were Mala-Hassan

expressed highest inhibition rate 34.417% and both

varieties of Baetmuni and Sarqala-Bakrajo recorded

only 3.333% inhibition of germination rate. The variety

of Sarqala-Bakrajo has highest speed of germination

when recorded 12.009 seed/ day and the variety of

Shekh-nuraddin recorded the lowest speed of

germination 6.245 seed/day. While parameters of

radical length, radical elongation velocity, plumule

length and plumule elongation velocity highest values

were observed with the variety of Sarqala-bakrajo but

the lowest values were associated with three varieties

Taefi, Mala-Hassan and Toli-Mawelian.

73 Ali and Maulood

Int. J. Biosci. 2011

Table 1. Effect of Grapevine variety on germination and some growth parameters of lettuce seeds.

Variety Germination Rate

Inhibition Rate

Germination Speed

(seed/day)

Radical length (cm)

Radical Speed of

elongation (cm/day)

Plumule length (cm)

Plumule Speed of

elongation (cm/day)

Taefi 77.083 c 22.917 c 8.960 d 0.820 d 0.103 d 0.623 d 0.078 c

Zareg-dohuok 84.583 b 15.417 d 7.399 e 1.088 c 0.137 c 0.720 cd 0.093 c

Baetmuni 96.667 a 3.333 e 10.654 c 1.929 b 0.242 b 1.092 b 0.160 b

Black Manga 92.083 a 7.917 e 11.051 bc 1.236 c 0.156 c 0.858 c 0.107 bc

Sarqala-Bakrajo 96.667 a 3.333 e 12.009 a 2.295 a 0.289 a 1.283 a 0.233 a

Awelka 93.750 a 6.250 e 9.572 d 1.733 b 0.218 b 1.117 b 0.137 bc

Rash merii 95.417 a 4.583 e 11.623 ab 1.303 c 0.164 c 0.822 c 0.103 bc

Mala Hassan 64.583 e 34.417 a 7.756 e 0.820 d 0.103 d 0.623 d 0.078 c

Ashgar Basraha 67.917 de 32.083 ab 9.622 d 1.160 c 0.146 c 0.802 c 0.100 bc

Kamalii 76.667 c 23.333 c 9.396 d 1.077 c 0.136 c 0.747 cd 0.093 c

Toli Mawelian 75.000 cd 25.000 bc 7.637 e 0.820 d 0.103 d 0.623 d 0.078 c

Shekh nuraddin 68.333 de 31.667 ab 6.245 f 1.220 c 0.153 c 0.822 c 0.103 bc

Common letter means that there was non-significant at 1% probability level by Duncan's test.

Table 2. The effect of Grapevine berries aqueous extracts concentration on some studied parameters.

Concentration Germination Rate

Inhibition Rate

Germination Speed

(seed/day)

Radical length (cm)

Radical Speed of

elongation (cm/day)

Plumule length (cm)

Plumule Speed of

elongation (cm/day)

control 100.00 a 0.000 c 13.663 a 3.280 a 0.413 a 2.493 a 0.312 a

5 % 85.139 b 14.861 b 9.106 b 1.135 b 0.143 b 0.523 b 0.097 b

10 % 80.9722 b 19.028 b 8.679 b 0.401 c 0.050 c 0.229 c 0.029 c

15 % 63.472 c 36.528 a 5.859 c 0.350 c 0.044 c 0.131 d 0.017 c

Common letter means that there was non-significant at 1% probability level by Duncan's test.

Aweilka variety has highest radical, plumule and total

seedling dry weights 14.542mg, 9.767mg and

24.309mg respectively, meanwhile the three varieties

of Taefi, Mala-Hassan and Toli-Mawelian recorded

lowest values of radical, plumule and total seedling dry

weights which were 11.667mg 8.367mg and 20.034mg

respectively (figure-1). Radical and plumule growth

inhibition rate were at highest levels with the three

varieties of Taefi, Mala-Hassan and Toli-Mawelian

which was 75% while the lowest record data were

29.811% of radical growth and 48.489% of plumule

growth (Fig. 2).

These results revealed the existence of allelopathic

compounds in the aqueous extracts of grapevine

berries with differences of the these allelochemicals

effect of each grapevine variety which causes the

inhibitory effects on the seed germination rate and the

growth of the seedling of lettuce seeds (Khan et

al.,1999, Akhtar et al.,2001 and Uremis et al., 2005),

74 Ali and Maulood

Int. J. Biosci. 2011

Table 3. Effect of the combination of grapevaline variety and aqueous extracts concentration on germination and

growth parameters of lettuce seeds.

The effect of Grapevine berries aqueous extracts

concentration on the studies parameters

Table (2) illustrates the significant effect (P≥0.01) of

different concentrations of grapevine berries aqueous

extracts on some of the data that was recorded in this

study. The highest values for germination rate 100%,

speed of germination 13.663 seed.day-1, radical length

3.280cm, radical elongation velocity 0.413cm.day-1,

plumule length 2.493cm, plumule elongation velocity

0.312 cm.day-1, were observed with the control

75 Ali and Maulood

Int. J. Biosci. 2011

treatments while lowest data for the above parameters

were recorded with the highest studied concentration

which was15% of grapevine berries aqueous extracts,

mean while the germination inhibition rate studied

highest data 36.528% was recorded with concentration

15%. Lettuce seedlings plumule, radical and total dry

weight 33.466mg, 46.667mg and 80.133mg

respectively were recorded with control treatments

mean while lowest data 0.311 mg, 1.072 mg and

1.383mg were observed at highest studied

concentration 15% (Figure-3). Plumule and radical

growth inhibition rates recorded highest values

94.704% and 89.283% respectively were allocated with

the highest studies concentrations 15% of grapevine

berries aqueous extracts meanwhile the lowest data

were recorded with control treatments (Fig. 4).

Table 4. Effect of the combination of grapevaline variety and aqueous extracts concentration on some growth

parameters of lettuce seeds.

76 Ali and Maulood

Int. J. Biosci. 2011

Fig. 1. The effect of grapevine varieties on the lettuce seedlings plumule, radical and total seedlings dry weight (mg).

Results of this study suggest that plant species

response will be differ to different concentrations of

the used aqueous extract (Rafiqul-Hoque et al., 2003

and Ben hammuda et al., 2001) which be an ideal

evidence of the concentration effect of the compounds

that exist in the used aqueous extracts (Ussalam et al.,

2011).

Table 5. Grapevine varitey chemical characters.

Variety Glycoside Saponines Tannins Phenolic compounds

Alkaloids

Taefi + - + + -

Zareg-dohuok + - + + -

Baetmuni + - + + -

Black-Manga + - + + -

Sarqala-Bakrajo + - + + +

Awelka + - + + -

Rash-merii + - + + +

Mala-Hassan + - + + +

Ashgar-Basraha + - + + -

Kamalii + - + + +

Toli-Mawelian + - + + -

Shekh-nuraddin + - + + +

77 Ali and Maulood

Int. J. Biosci. 2011

Fig. 2. The effect of grapevine varieties on Lettuce

seedlings plumule and radical growth inhibition rates.

Fig. 3. The effect of grapevine berries aqueous extracts

on lettuce seedlings plumule, radical and total dry

weights (mg).

The effect of combination of grapevine varieties and

grapevine berries aqueous extracts concentration on

the studies parameters

The combination of the two studied factors grapevine

varieties and grapevine berries aqueous extracts

revealed significant differences (P≥0.01) on

germination rate, inhibition of germination,

germination speed, plumule and radical length,

elongation speed of both plumule and radical of lettuce

seedlings (Table 3). Whereas highest germination rate

100% was recorded with control treatments of all

studied varieties, meanwhile highest inhibition of

germination rate was 68.333 % recorded with the

concentration of 15% of grapevine berries of Shek-

nuraddin variety. Highest germination speed was

13.633 seedling.day-1 considered with control

treatment, mean while lowest germination speed

revealed 1.420 seed.day-1 with concentration of 15% of

Mala-hassan variety, highest radical and plumule

length were observed with control treatment and the

same phenomenon was noticed with radical and

plumule elongation speed. Lettuce seedlings plumule

and radical dry weight were observed recorded and

obtained significant differences as shown in (Table 4).

Fig. 4. The effect of grapevine Berries aqueous

extracts concentration on Lettuce seedlings plumule

and radical growth inhibition rates.

Fig. 5. The levels of total polyphenols in the grapevine

berries in part per million.

Grapevine berries chemical characters

The existence of chemical compounds such as

Alkaloids, poly phenolic compounds, Tannins,

Saponnins, and Glycosids for the twelve grapevine

varieties were recorded as shown in (Table 5), where

there is obvious differences in the existence of

alkaloids which only exist in five verities, meanwhile

poly phenolic compounds, tannins and Saponnins exist

in all studied varieties. Polyphenolic compounds were

at highest level in Black-manga variety (figure-5), while

total soluble compounds were at highest level in

ashgar-Basrah variety (Fig. 6).

78 Ali and Maulood

Int. J. Biosci. 2011

Fig. 6. The total soluble solids compound percentages

in grapevine berries.

In this study grapevine berries exposed different levels

of total soluble solid compounds and total poly phenol

levels which may reveals the inhibitory effects of the

aqueous extracts, mean while increasing the

concentration lead to rising the inhibitory effect in all

tested grapevine varieties (Taiz and Zeiger, 2006;

Lambers et al., 2008).

Conclusion

This research demonstrates the allelopathical potential

of grapevine berries aqueous extracts which differs

from variety to another and the inhibitory effect

increased with increasing the concentration the tested

aqueous extract. These inhibitory effects may be

related to the different compound that needs more

studies to reveal the responsible compounds.

Acknowledgments

We would like to thank (Erbil research center-

Kurdistan region-Iraq) for their assistance and help

during the implementation of these experiments.

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